LOCUS       sequence-471891-        4779 bp    DNA     circular SYN 26-JUN-2025
DEFINITION  Express Max-SmBiT Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4779)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBiT Assay Vector, Max
FEATURES             Location/Qualifiers
     source          1..4779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1458..1469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1722..1740)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1777..1898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1788..1807)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1842..1861
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2143..2500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2143..2334
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2166..2186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2351..2486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2515..2562
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2594..3388
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2648..2667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3258..3277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3452..3500
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3739..4327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3819..3838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4443..4726
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgagc gataacgatg
     1081 acatcgaggt ggagagcgac gaagagcaac cgaggtttca atctgcggct gacaaacggg
     1141 ctcatcataa tgcactggaa cgaaaacgta gggaccacat caaagacagc tttcacagtt
     1201 tgcgggactc agtcccatca ctccaaggag agaaggcatc ccgggcccaa atcctagaca
     1261 aagccacaga atatatccag tatatgcgaa ggaaaaacca cacacaccag caagatattg
     1321 acgacctcaa gcggcagaat gctcttctgg agcagcaagt ccgtgcactg gagaaggcga
     1381 ggtcaagtgc ccaactgcag accaactacc cctcctcaga caacagcctc tacaccaacg
     1441 ccaagggcag caccatctct gccttcgatg ggggctcgga ctccagctcg gagtctgagc
     1501 ctgaagagcc ccaaagcagg aagaagctcc ggatggaggc cagcgtttct caaggcagtt
     1561 caggtggtgg cgggagcgga ggtggaggct cgagcggtgt gaccggctac cggctgttcg
     1621 aggagattct gtaaggccgc gactctagag tcgacctgca ggcatgcaag ctgatccggc
     1681 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1741 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     1801 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1861 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1921 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     1981 aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2041 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     2101 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     2161 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2221 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2281 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2341 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2401 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2461 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac
     2521 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     2581 caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2641 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2701 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2761 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2821 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2881 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2941 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3001 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3061 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3121 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3181 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3241 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3301 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3361 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3421 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     3481 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     3541 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     3601 ggctttctac ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4141 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4321 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4381 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     4441 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
     4501 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
     4561 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
     4621 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
     4681 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
     4741 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
